PUMA publications for /user/bastian/Kinetics,https://puma.ub.uni-stuttgart.de/user/bastian/Kinetics,PUMA RSS feed for /user/bastian/Kinetics,2024-03-28T21:07:59+01:00Acetohydroxyacid synthase, a novel target for improvement of L-lysine production by Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2ececc5edfb975851e24e89f723cab58c/bastianbastian2018-02-09T13:18:17+01:00Acetolactate Acid, Bacterial Butyrates Corynebacterium Deletion, Enzyme Expression Gene Inhibitors, Isoleucine, Kinetics, Leucine, Lysine, Profiling, Proteins, Pyruvic Sequence Synthase, Valine, glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stephan Hans" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/1"><span itemprop="name">S. Hans</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Brigitte Bathe" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/2"><span itemprop="name">B. Bathe</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/3"><span itemprop="name">B. Eikmanns</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Appl. Environ. Microbiol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">75 </span></span>(<span itemprop="issueNumber">2</span>):
<span itemprop="pagination">419--427</span></em> </span>(<em><span>January 2009<meta content="January 2009" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.jan2419--427Acetohydroxyacid synthase, a novel target for improvement of {L}-lysine production by {Corynebacterium} glutamicum752009Acetolactate Acid, Bacterial Butyrates Corynebacterium Deletion, Enzyme Expression Gene Inhibitors, Isoleucine, Kinetics, Leucine, Lysine, Profiling, Proteins, Pyruvic Sequence Synthase, Valine, glutamicum, myown The influence of acetohydroxy acid synthase (AHAS) on L-lysine production by Corynebacterium glutamicum was investigated. An AHAS with a deleted C-terminal domain in the regulatory subunit IlvN was engineered by truncating the ilvN gene. Compared to the wild-type AHAS, the newly constructed enzyme showed altered kinetic properties, i.e., (i) an about twofold-lower K(m) for the substrate pyruvate and an about fourfold-lower V(max); (ii) a slightly increased K(m) for the substrate alpha-ketobutyrate with an about twofold-lower V(max); and (iii) insensitivity against the inhibitors L-valine, L-isoleucine, and L-leucine (10 mM each). Introduction of the modified AHAS into the L-lysine producers C. glutamicum DM1729 and DM1933 increased L-lysine formation by 43\% (30 mM versus 21 mM) and 36\% (51 mM versus 37 mM), respectively, suggesting that decreased AHAS activity is linked to increased L-lysine formation. Complete inactivation of the AHAS in C. glutamicum DM1729 and DM1933 by deletion of the ilvB gene, encoding the catalytic subunit of AHAS, led to L-valine, L-isoleucine, and L-leucine auxotrophy and to further-improved L-lysine production. In batch fermentations, C. glutamicum DM1729 Delta ilvB produced about 85\% more L-lysine (70 mM versus 38 mM) and showed an 85\%-higher substrate-specific product yield (0.180 versus 0.098 mol C/mol C) than C. glutamicum DM1729. Comparative transcriptome analysis of C. glutamicum DM1729 and C. glutamicum DM1729 Delta ilvB indicated transcriptional differences for about 50 genes, although not for those encoding enzymes involved in the L-lysine biosynthetic pathway.Effect of pyruvate dehydrogenase complex deficiency on L-lysine production with Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2930ecfe113b73f241ab0452b05337d89/bastianbastian2018-02-09T13:18:17+01:00Bacterial, Base Biotechnology Complex, Corynebacterium DNA, Dehydrogenase Deletion, Expression, Fermentation, Gene Genes, Kinetics, Lysine, Pyruvate Sequence, glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/197097cd65b8e231a2578efbc7586a5dc/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Mark E. Schreiner" itemprop="url" href="/person/197097cd65b8e231a2578efbc7586a5dc/author/1"><span itemprop="name">M. Schreiner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Matthias Moch" itemprop="url" href="/person/197097cd65b8e231a2578efbc7586a5dc/author/2"><span itemprop="name">M. Moch</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Marco Oldiges" itemprop="url" href="/person/197097cd65b8e231a2578efbc7586a5dc/author/3"><span itemprop="name">M. Oldiges</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/197097cd65b8e231a2578efbc7586a5dc/author/4"><span itemprop="name">B. Eikmanns</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Appl. Microbiol. Biotechnol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">76 </span></span>(<span itemprop="issueNumber">3</span>):
<span itemprop="pagination">615--623</span></em> </span>(<em><span>September 2007<meta content="September 2007" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Microbiol. Biotechnol.sep3615--623Effect of pyruvate dehydrogenase complex deficiency on {L}-lysine production with {Corynebacterium} glutamicum762007Bacterial, Base Biotechnology Complex, Corynebacterium DNA, Dehydrogenase Deletion, Expression, Fermentation, Gene Genes, Kinetics, Lysine, Pyruvate Sequence, glutamicum, myown Intracellular precursor supply is a critical factor for amino acid productivity of Corynebacterium glutamicum. To test for the effect of improved pyruvate availability on L-lysine production, we deleted the aceE gene encoding the E1p enzyme of the pyruvate dehydrogenase complex (PDHC) in the L-lysine-producer C. glutamicum DM1729 and characterised the resulting strain DM1729-BB1 for growth and L-lysine production. Compared to the host strain, C. glutamicum DM1729-BB1 showed no PDHC activity, was acetate auxotrophic and, after complete consumption of the available carbon sources glucose and acetate, showed a more than 50\% lower substrate-specific biomass yield (0.14 vs 0.33 mol C/mol C), an about fourfold higher biomass-specific L-lysine yield (5.27 vs 1.23 mmol/g cell dry weight) and a more than 40\% higher substrate-specific L-lysine yield (0.13 vs 0.09 mol C/mol C). Overexpression of the pyruvate carboxylase or diaminopimelate dehydrogenase genes in C. glutamicum DM1729-BB1 resulted in a further increase in the biomass-specific L-lysine yield by 6 and 56\%, respectively. In addition to L-lysine, significant amounts of pyruvate, L-alanine and L-valine were produced by C. glutamicum DM1729-BB1 and its derivatives, suggesting a surplus of precursor availability and a further potential to improve L-lysine production by engineering the L-lysine biosynthetic pathway.